Renin protein detection in Western blotting - (Sep/12/2007 )

I would really appreciate if someone out there could help me find my way out of this. I have done a few Westerns for renin extracted from mice kidney and was able to obtain some bands.

However, of recent , using the same protocols and same conditions ,I have struggled endlessly to reproduce the the results all to no avail.

Please can someone tell me what might be the problem as i have checked all my reagents, even made new stock and still got nothing. Please HELP!!!

-ojbillions-

What control do you use and how is that performing?

We measure renin activity rather than the mass. We've got to be so careful with sample collection and stroreage because of the prorenin activation issue. That produces false positives, though. What conditions do you store your samples?

-paraboxa-

are you using the same sample? maybe the renin decomposed.

do you see any bands in the western (smaller (decomposed) or larger (aggregated))?